1.Human erythropoietin-modified human amniotic mesenchymal stem cellsvia subarachnoid transplantation promote neurologic recovery from brain injury
Chinese Journal of Tissue Engineering Research 2016;20(23):3432-3438
BACKGROUND:Studies have shown that as a regulator of bone marrow functionerythropoietinis a glycoprotein that controls the development of the central nervous system and has neurotrophic and neuroprotective potential. Therefore, transplantation of human amniotic mesenchymal stem cels geneticaly modified by human erythropoietin is a new choice for brain injury treatment.
OBJECTIVE:To observe the effect of transplantation of human amniotic mesenchymal stem cels geneticaly modified byhuman erythropoietin on the functional recovery from brain injury in rats.
METHODS:Eukaryotic expression plasmid pcDNA3.1 carrying erythropoietin was successfuly constructed and transferred into amniotic mesenchymal stem cels culturedin vitro. Expression of erythropoietin was detected using western blot assay before and after transfection. Rat models of middle cerebral arterial occlusion was made and given transplantation of transfected amniotic mesenchymal stem celsviathe tail vein (transfection group). Additionaly, model and simple cel transplantation groups were set in a comparative study.
RESULTS AND CONCLUSION:Findings from western blot detection showed that transfected cels could express human erythropoietin. Compared with the other groups, modified neurologic severity scores, growth-associated protein 43 and aquaporin 9 at mRNA and protein levels were al decreased significantly in the transfection group. Furthermore, the number of cels positive for CM-Dil was highest in the transfectiongroup, folowed by simple cel transplantation group, and lowest in the model group (alP<0.05). Overal findings from this study show that human erythropoietin-modified human amniotic mesenchymal stem cel transplantation promotes neurologic recoveryfrom brain injury through eliciting a reduction in growth-associated protein 43 and aquaporin 9 at mRNA and protein levels as wel as inhibiting cel apoptosis.
2.Tissue engineered artificial nerves repair sciatic nerve defect in rats:an evaluation using horseradish peroxidase retrograde tracer technique
Chinese Journal of Tissue Engineering Research 2014;(29):4658-4662
BACKGROUND:We have previously prepared acellular nerve graft and implanted bone marrow mesenchymal stem cells into the graft, to successful y construct tissue engineered artificial nerves.
OBJECTIVE:Horseradish peroxidase nerve retrograde tracer technique was used to evaluate protective effects on sensory neurons fol owing sciatic nerve defect bridging with tissue engineered artificial nerves constructed by acellular nerve graft and bone marrow mesenchymal stem cells in rats.
METHODS:Adult, clean, healthy, male Sprague-Dawley rats were randomly assigned to three group:(1) Experimental group:Rat sciatic nerve detect was bridged by acellular nerve graft combined with bone marrow mesenchymal stem cells;(2) Blank control group:Rat sciatic nerve defect was bridged by acellular nerve graft;(3) Autologous nerve control group:Rat sciatic nerve defect was bridged by autologous nerve transplantation. Regeneration of sensory neurons in the spinal dorsal root ganglia was assessed using horseradish peroxidase nerve retrograde tracer technique at 12 weeks fol owing surgery.
RESULTS AND CONCLUSION:Sensory neuron regeneration in the spinal dorsal root ganglia at 12 weeks fol owing surgery was better in the experimental group compared with blank control group. No significant difference was detected between experimental group and autologous nerve control group. S-100 immunohistochemical staining in plantar skin showed brown positive reaction in each group. These findings indicate that tissue engineered artificial nerves constructed by acellular nerve graft and bone marrow mesenchymal stem cells have protective effects on sensory neurons in the spinal dorsal root ganglia, and can promote the recovery of sensory function and repair sciatic nerve defect in rats.
3.Efficacy of Qiju Dihuang pills oral combined with polyethylene glycol external in meibomian gland dysfunction dry eye syndrome
Chinese Journal of Biochemical Pharmaceutics 2016;36(6):139-141
Objective To investigate the clinical efficacy of Qiju Dihuang pills oral combined with polyethylene glycol external in the treatment of meibomian gland dysfunction dry eye syndrome.Methods A total of 144 cases with meibomian gland dysfunction dry eye syndrome, a total of 288 eyes, were collected and divided into two groups with 72 patients in each group.Patients in the control group were treated by polyethylene glycol external, and patients in the experimental group were treated by Qijudihuang pills oral combined with polyethylene glycol external.The tear film break-up time ( BUT) , schirmer test (schimer-1) and corneal fluorescent (FL) and dry eye subjective symptoms were recorded before treatment and after treatment 2 and 4 weeks,at the same time the clinical efficacy was compared.Results The clinical effective rate of the control group was 82.64%, which was lower than the experimental group (93.06%) after treatment, with statistical significance (P<0.05); compared with before treatment, the tear film BUT and schirmer test of the experimental group were higher than the control group after treatment 2 and 4 weeks, corneal FL and subjective symptoms scored were lower than the control group, with statistical significance (P<0.05).Conclusion Qiju Dihuang pills oral combined with polyethylene glycol external could effectively release the meibomian gland dysfunction dry eye symptoms and the effect is obvious.
4.Expression and biological effects of pcDNA_(3.1)-OGP gene transfection on rabbit bone marrow stromal cells in vitro
Gang AN ; Songcen LV ; Qiukui DENG
Orthopedic Journal of China 2006;0(03):-
[Objective]To observe the expression of osteogenic growth peptide(OGP) gene,which had been transfected into rabbit bone marrow stromal cells(BMSC) by adenovirus,and detect the proliferation and differentiation of rabbit bone marrow stromal cells after gene transfection in vitro.[Methods]pcDNA3.1-OGP was constructed by using gene clone and recombined technique.With the help of lipofectamine 2000,BMSC were transfected with pcDNA3.1-OGP.The positive cell clones were selected with G418.The expression of OGP gene was observed by RT-PCR.Alkaline phosphatase(ALP) activity and type I collagen in pcDNA3.1-OGP transfected bone marrow stromal cells were measured to observe the differentiation from bone marrow stromal cells into osteoblast lineage.[Results]The pcDNA3.1-OGP vector was constructed successful.According to analysis of RT-PCR,alkaline phosphatase(ALP) and type I collagen assay showed that OGP gene could be expressed in BMSC,and could induce bone marrow stromal cells differentiation into osteoblast lineage.[Conclusion]The pcDNA3.1-OGP vector can be constructed successfully and its expression is confirmed in BMSC.The expression of OGP in bone marrow stromal cells can induce BMSC differentiation into osteoblast lineage.
6.Effects of Morinda on receptor activator of nuclear factor-kappaB expression in osteoclasts in osteoporosis
Desheng MIAO ; Gang LV ; Xinyuan MO
Chinese Journal of Tissue Engineering Research 2016;20(24):3516-3522
BACKGROUND:Morinda has been reported to promote the proliferation, the secretion of alkaline phosphatase and osteocalcin, and mRNA expression of transforming growth factor of osteoblasts. However, little information is available addressing the effects of Morinda on receptor activator of nuclear factor-κB expression in osteoclasts in rats with osteoporosis. OBJECTIVE:To study the effects of Morinda on receptor activator of nuclear factor-κB expression in osteoclastsofosteoporosis rats. METHODS:Thirty Sprague-Dawley rats were equaly and randomly divided into Morinda and 17β-estradiol groups. Rat models of osteoporosis were established by bilateral ovariectomy, and then 5 mL of Morinda decocta(1.0mmol/L)and 17β-estradiol (1×10-6mmol/L) were administered intragastricaly to rats in Morinda and 17β-estradiol groups for 3 consecutive months, respectively. Primary osteoclasts were isolated from rats in both groups, andthen cultured for 3, 6 and 9 days folowed by TRAP staining andcelcounting. Bone mineral density of the proximal and distal femur, urine and serum levels of Ca2+and progesterone, and receptor activator of nuclear factor-κB expression in osteoclasts ofrats in both groups were determined. RESULTS AND CONCLUSION:Osteoclast fusion was reduced in Morinda group. In contrast, number of osteoclastswas increased andcels becamemore maturein the17β-estradiol group. Bone mineral density of the proximal and distal femur bilateraly, urine and serum levels of Ca2+and progesterone were significantly increased, while receptor activator of nuclear factor-κB expression was significantly decreased in osteoclasts in Morinda group compared with 17β-estradiol group (P< 0.05). These results indicate that Morinda reduces receptor activator of nuclear factor-κB expression in osteoclasts in osteoporosis rats, thereby inhibiting the development and progression of osteoporosis.
7.Chitosan conduits with human umbilical cord mesenchymal stem cells induce differentiation and growth of the nerve lateral bud
Xuepu ZHANG ; Qiang XIAO ; Gang LV
Chinese Journal of Tissue Engineering Research 2014;(43):6985-6989
BACKGROUND:Human umbilical cord mesenchymal stem cells are similar to bone marrow mesenchymal stem cells, which can directional y differentiate into neuron-like cells, secrete various cytokines, and provide the base for nerve regeneration. <br> OBJECTIVE:To study the role of chitosan composite nerve conduit carrying umbilical cord mesenchymal stem cells in nerve end-to-side anastomosis. <br> METHODS:Thirty while rabbits were randomized into three groups. The central branch of the right posterior peroneal nerve were cut and proximal y ligated, and then sutured evaginably to the muscle. In the control group, the distal end of the common peroneal nerve were anastomosed into the tibial nerve at 30°-45°;in the stenting group, the chitosan conduit was bridged at the same interval and angle into the end-to-side anastomosis site between the tibial nerve and peroneal nerve;in the cel-stenting group, the chitosan conduit carrying human umbilical cord mesenchymal stem cells was bridged at the same interval and angle into the end-to-side anastomosis site between the tibial nerve and peroneal nerve. After 12 weeks, gross observation, neurophysiological examination and anti-S-100 immunohistochemistry detection were performed. <br> RESULTS AND CONCLUSION:After 12 weeks of operation, in the cel-stenting group, the conduit degraded completely, the nerve diameter was similar to that of the normal peroneal nerve, and the motor nerve conduction velocity was higher than that in the control and stenting groups (P<0.01). Anti-S-100 immunohistochemistry results showed that a great amount of brownish red Schwann cells arranged around the regenerated nerve fibers in the cel-stenting group, while there was few and sparse brownish red substance, and the Schwann cells grew worse in the stenting and control groups. These findings suggest that umbilical cord mesenchymal stem cells have an obvious role in promoting nerve regeneration, induce bud growth, accelerate the growth rate of regenerated fibers, and improve growth and maturity of Schwann cells.
8.Expression and pathological significance of CD-147 protein in osteosarcoma
Qiang LU ; Gang LV ; Yanfeng WANG
Orthopedic Journal of China 2006;0(07):-
[Objective]To investigate the expressions of CD-147 in osteosarcoma samples and to research clinically pathological significance. [Methods]The expressions of CD-147 was detected by immunohistochemistry SABC method in 55 cases of osteosarcoma samples.[Results]The moderate/strong expression of CD-147 was 54.5% compared with giant cell tumor of bone.And the high expressions of CD-147 was in close correlation with diagnosis and prognosis in osteosarcoma.[Conclusion]CD-147 were overexpressed in osteosarcoma,which can be relevant to the malignant progression of osteosarcoma.
9.Expeiment study on the proliferation and plasticity of ependymal cells in situ in the injuried spinal cord of adult rats
Ying CAO ; Gang LV ; Yanfeng WANG
Orthopedic Journal of China 2006;0(18):-
[Objective]To observe the proliferation and plasticity of neural stem cells in sim in adult rats after spinal cord injury.[Method]Spinal cord injury models were made in 60 wistar rats and the dynamic expression of bromodeoxyuridine(BrDU) and polysialylated ependymal cells adhesion molecule(PSA-NCAM) were determined by immuno-histochemisty.[Result]Compared with the controls,the number of Brdupositive cells in the injured spinal cord increased strikingly on the 1 st day (P
10.Study on Methods for Population Size Estimation of HIV High Risk Groups and its Application in HIV/AIDS Situation Analysis in China
Peng XU ; Gang ZENG ; Fan LV ;
Journal of Medical Research 2006;0(05):-
Objective To investigate HIV high risk population size estimation method,analyze the epidemic situation in China and offer scientific evidence for policy making. Methods Appropriate methods to estimate different type high risk populations were ascertained by experimental study. Results Size estimation methods and procedures for different type high risk population were formed.Epidemic estimation and parameters fit for China were ascertained.HIV epidemic situation and characteristics were stated. Conclusion Methods formed in the study are the key points for the understanding of number of PLWHA,for the exact judgment of HIV epidemic,for the improvement of inspection system and examination work,for the formation of "Four Free and One Care".